Abstract

Low back pain and disc degeneration affect quality of life and imposes an enormous financial burden. Although annulus fibrosus (AF) tissue engineering provides an alternative therapeutic possibility in the treatment of degenerative intervertebral disc disease, it is restricted by the biochemical properties, organizational structure, and mechanical characteristics of the scaffold. The ideal scaffold should closely mimic the natural extracellular matrix (ECM) in structure and function for long-term stability and survival. Poly(ether carbonate urethane) urea (PECUU) can be electrospun into nanofibrous scaffolds to mimic ECM architecture with the appropriate mechanical properties. However, PECUU scaffolds lack the bioactivity of natural ECM. On the other hand, a decellularized annulus fibrosus matrix (DAFM) has good biocompatibility and biodegradability and has been shown to promote secretion of AF-related ECM. Herein, DAFM/PECUU-blended electrospun scaffolds were fabricated with the help of coaxial electrospinning technology for the first time. AF-derived stem cells were cultured on DAFM/PECUU electrospun scaffolds, and cellular metabolic activity, morphology, and gene expression assays as well as AF-related ECM synthesis were performed. The results showed that AF-derived stem cells proliferated well on the scaffolds. Gene expression and ECM secretion of collagen type I and II and aggrecan from AF-derived stem cells cultured on DAFM/PECUU electrospun scaffolds were higher than from those on PECUU fibrous scaffolds. Thus, DAFM/PECUU electrospun scaffolds are a potential candidate for AF tissue engineering applications.

摘要

腰痛和椎间盘退变会影响生活质量并带来巨大的经济负担。尽管纤维环（AF）组织工程在退行性椎间盘疾病的治疗中提供了另一种治疗可能性，但它受到支架的生化特性、组织结构和机械特性的限制。理想的支架应在结构和功能上密切模仿天然细胞外基质 (ECM)，以实现长期稳定性和存活。聚（醚碳酸酯氨基甲酸酯）尿素（PECUU）可以电纺成纳米纤维支架，以模拟具有适当机械性能的 ECM 结构。然而，PECUU 支架缺乏天然 ECM 的生物活性。另一方面，脱细胞纤维环基质 (DAFM) 具有良好的生物相容性和生物降解性，已被证明可促进 AF 相关 ECM 的分泌。在此，首次借助同轴静电纺丝技术制造了 DAFM/PECUU 混合的静电纺丝支架。AF 衍生的干细胞在 DAFM/PECUU 电纺支架上培养，并进行细胞代谢活性、形态和基因表达测定以及 AF 相关 ECM 合成。结果表明，AF 来源的干细胞在支架上增殖良好。在 DAFM/PECUU 电纺支架上培养的 AF 衍生干细胞的 I 型和 II 型胶原蛋白和蛋白聚糖的基因表达和 ECM 分泌高于 PECUU 纤维支架上的那些。因此，